6-aminoisoquinoline compounds

ABSTRACT

6-Amino isoquinoline compounds are provided that influence, inhibit or reduce the action of a kinase. Pharmaceutical compositions including therapeutically effective amounts of the 6-aminoisoquinoline compounds and pharmaceutically acceptable carriers are also provided. Various methods using the compounds and/or compositions to affect disease states or conditions such as cancer, obesity and glaucoma are also provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of prior application Ser. No.14/583,439 filed Dec. 26, 2014, which is a continuation of priorapplication Ser. No. 13/723,811, filed Dec. 21, 2012, which is acontinuation of prior application Ser. No. 13/017,708, filed Jan. 31,2011, which is a continuation of prior application Ser. No. 11/621,887,filed Jan. 10, 2007, each of which is incorporated herein by referencein its entirety.

FIELD OF THE INVENTION

The present invention relates to 6-aminoisoquinoline compounds thataffect the function of kinases in a cell and that are useful astherapeutic agents or with therapeutic agents. In particular, thesecompounds are useful in the treatment of eye diseases such as glaucomaand for diseases characterized by abnormal growth, such as cancers.

BACKGROUND

A variety of hormones, neurotransmitters and biologically activesubstances control, regulate or adjust the functions of living bodiesvia specific receptors located in cell membranes. Many of thesereceptors mediate the transmission of intracellular signals byactivating guanine nucleotide-binding proteins (G proteins) to which thereceptor is coupled. Such receptors are generically referred to asG-protein coupled receptors (GPCRs) and include, among others,α-adrenergic receptors, β-adrenergic receptors, opioid receptors,cannabinoid receptors and prostaglandin receptors. The effect of thesereceptors is not direct but mediated by a host of intracellularproteins. The importance of these secondary, or “downstream” proteins isonly now being recognized and investigated as potential interventionpoints in disease states. One of the most important classes of thesedownstream proteins is the “kinase” class.

The various kinases play an important role in the regulation of variousphysiological functions. By way of example, kinases have been implicatedin a number of disease states, including, but not limited to: cardiacindications such as angina pectoris, essential hypertension, myocardialinfarction, supraventricular and ventricular arrhythmias, congestiveheart failure, atherosclerosis, renal failure, diabetes, respiratoryindications such as asthma, chronic bronchitis, bronchospasm, emphysema,airway obstruction, upper respiratory indications such as rhinitis,seasonal allergies, inflammatory disease, inflammation in response toinjury, rheumatoid arthritis. The importance of p38 MAPK inhibitors asnew drugs for rheumatoid arthritis is reflected by the large number ofcompounds that has been developed over the last years (J. Westra and P.C. Limburg Mini-Reviews in Medicinal Chemistry Volume 6, Number 8,August 2006) Other conditions include chronic inflammatory boweldisease, glaucoma, hypergastrinemia, gastrointestinal indications suchas acid/peptic disorder, erosive esophagitis, gastrointestinalhypersecretion, mastocytosis, gastrointestinal reflux, peptic ulcer,Zollinger-Ellison syndrome, pain, obesity, bulimia nervosa, depression,obsessive-compulsive disorder, organ malformations (for example, cardiacmalformations), neurodegenerative diseases such as Parkinson's Diseaseand Alzheimer's Disease, multiple sclerosis, Epstein-Barr infection andcancer (Nature Reviews Drug Discovery 1, 493-502 2002). In other diseasestates, the role of kinases is only now becoming clear. The retina is acomplex tissue composed of multiple interconnected cell layers, highlyspecialized for transforming light and color into electrical signalsperceived by the brain. Damage or death of the primary light-sensingcells, the photoreceptors, results in devastating effects on vision.Despite the identification of numerous mutations that cause inheritedretinal degenerations, the cellular and molecular mechanisms leadingfrom the primary mutations to photoreceptor apoptosis are not wellunderstood, but may involve the wnt pathway (AS Hackam The Wnt SignalingPathway in Retinal Degeneration IUBMB Life Volume 57, Number 6/June2005).

The success of the tyrosine-kinase inhibitor STI571 (Gleevec) in thetreatment of chronic myelogenous leukaemia (Nature Reviews DrugDiscovery 2, 296-313 2003) has spurred considerable efforts to developother kinase inhibitors for the treatment of a wide range of othercancers (Nature Reviews Cancer 3, 650-665 2003). The balance between theinitiation and the inactivation of the intracellular signals regulatesthe intensity and duration of the response of the receptors to stimulisuch as agonists. When desensitization occurs, the mediation orregulation of the physiological function mediated or regulated by the Gproteins to which the receptors are coupled is reduced or prevented. Forexample, when agonists are administered to treat a disease or conditionby activation of certain receptors, the receptors become desensitizedfrom the action of the GRKs such that agonist administration may nolonger result in therapeutic activation of the appropriate receptors. Atthat point, administration of the agonist no longer enables sufficientor effective control of or influence on the disease or conditionintended to be treated.

In view of the role of kinases in many disease states, there is anurgent and continuing need for ligands which inhibit or modulate theactivity of kinases. Without wishing to be bound by theory, it isthought that modulation of the activity of kinases by the compounds ofthe present invention is responsible for their beneficial effects.

SUMMARY

In a first aspect of the invention, a compound is provided according toFormula (I):

wherein A is —CH₂NH—,

—SCH₂—, —CH₂S(O)—, —CH₂S(O)(O)—, —S(O)CH₂—, —S(O)(O)CH₂—, —CH₂CH₂—,—CH(R₁₀)CH₂—, —CH₂CH(R₁₀), —CH═CH—,

wherein R₁₀ is hydrogen, unsubstituted C₁-C₄ alkyl, C₁-C₄ alkenyl, C₁-C₄alkynyl, or amino;

wherein R₁, and R₂ are, independently, hydrogen, hydroxyl, halogen,C₁-C₄ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, amino, nitro, cyano, C₁-C₄carbonyl, C₁-C₄ carbonylamino, C₁-C₄ alkoxy, C₁-C₄ sulfonyl, C₁-C₄sulfonylamino, C₁-C₄ thioalkyl or C₁-C₄ carboxyl; and

wherein R₃ is hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, amino,cyano, cycloalkyl, heterocycloalkyl, aryl, C₁-C₄ alkyl aryl, heteroaryl,C₁-C₄ alkyl heteroaryl, carbonyl, carbonylamino, thioalkyl, sulfonyl,sulfonylamino, acyl, or carboxyl.

In a second aspect, a compound is provided according to Formula II:

wherein R₁ and R₂ are, independently, hydrogen, hydroxyl, halogen, C₁-C₄alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, amino, nitro, cyano, C₁-C₄carbonyl, C₁-C₄ carbonylamino, C₁-C₄ alkoxy, C₁-C₄ sulfonyl, C₁-C₄sulfonylamino, C₁-C₄ thioalkyl or C₁-C₄ carboxyl;

wherein R₄ is hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, amino,cyano, cycloalkyl, heterocycloalkyl, aryl, C₁-C₄ alkyl aryl, heteroaryl,C₁-C₄ alkyl heteroaryl, carbonyl, carbonylamino, thioalkyl, sulfonyl,sulfonylamino, acyl, or carboxyl; and

wherein B is a chain containing from 0 to 3 member atoms, X represents nindependently chosen member atoms which together form a ring structureand n is an integer from about 0 to about 5.

In another aspect, a compound is provided according to Formula III:

wherein one of X₁, X₂ and X₃ is independently selected from CH₂, O, S,S(O), S(O)(O),

and the other two of X₁, X₂ and X₃ are independently selected from CH₂,O, S, S(O), S(O)(O),

and bond;

wherein R₁, and R₂ are, independently, hydrogen, hydroxyl, halogen,C₁-C₄ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, amino, nitro, cyano, C₁-C₄carbonyl, C₁-C₄ carbonylamino, C₁-C₄ alkoxy, C₁-C₄ sulfonyl, C₁-C₄sulfonylamino, C₁-C₄ thioalkyl and C₁-C₄ carboxyl; and

wherein R₅ and R₁₀ are independently hydrogen, halogen, alkyl, alkenyl,alkynyl, alkoxy, amino, cyano, cycloalkyl, heterocycloalkyl, aryl, C₁-C₄alkyl aryl, heteroaryl, C₁-C₄ alkyl heteroaryl, carbonyl, carbonylamino,thioalkyl, sulfonyl, sulfonylamino, acyl, or carboxyl.

In a further aspect of the invention, a compound is provided accordingto Formula IV

wherein A is a substituted or unsubstituted linker consisting of atleast one member atom and at most 4 member atoms wherein the linker maybe mono- or disubstituted with halogen, cyano, nitro or C₁-C₄ alkyl, orthe substituted atoms may attach back to the main chain to form a ring;

wherein R₁, and R₂ are, independently, hydrogen, hydroxyl, halogen,C₁-C₄ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, amino, nitro, cyano, C₁-C₄carbonyl, C₁-C₄ carbonylamino, C₁-C₄ alkoxy, C₁-C₄ sulfonyl, C₁-C₄sulfonylamino, C₁-C₄ thioalkyl or C₁-C₄ carboxyl; and

wherein R₃ is hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, amino,cyano, cycloalkyl, heterocycloalkyl, aryl, C₁-C₄ alkyl aryl, heteroaryl,C₁-C₄ alkyl heteroaryl, carbonyl, carbonylamino, thioalkyl, sulfonyl,sulfonylamino, acyl, or carboxyl.

In yet a further aspect, a method is provided for influencing the actionof a kinase in a cell, a tissue, or a living mammal comprisingadministering to or contacting with the cell, tissue, or mammal at leastone compound according to claim 1, 6, 12 or 16, or increasing theeffectiveness of another therapeutic agent in a cell, tissue or livingmammal comprising administering to or contacting with the cell, tissueor mammal a therapeutically effective amount of at least one compoundaccording to claim 1, 6, 12 or 16.

In yet another aspect of the invention, a pharmaceutical composition isprovided, comprising:

a) a 6-aminoisoquinoline derivative having the structure

wherein R₁, and R₂ are, independently, hydrogen, hydroxyl, halogen,C₁-C₄ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, amino, nitro, cyano; C₁-C₄carbonyl, C₁-C₄ carbonylamino, C₁-C₄ alkoxy, C₁-C₄ sulfonyl, C₁-C₄sulfonylamino, C₁-C₄ thioalkyl or C₁-C₄ carboxyl; and

wherein R₄ is hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, amino,cyano, cycloalkyl, heterocycloalkyl, aryl, C₁-C₄ alkyl aryl, heteroaryl,C₁-C₄ alkyl heteroaryl, carbonyl, carbonylamino, thioalkyl, sulfonyl,sulfonylamino, acyl, or carboxyl; and

-   -   b) a carrier.

In yet a further aspect is provided a method of treating a conditioncomprising administering to a subject in need of treatment a safe andeffective amount of a 6-aminoisoquinoline derivative, wherein thecondition is selected from the group consisting of eye disease, bonedisorder, obesity, heart disease, hepatic disease, renal disease,pancreatitis, cancer, myocardial infarct, gastric disturbance,hypertension, fertility control, nasal congestion, neurogenic bladderdisorder, gastrointestinal disorder, and dermatological disorder.

DETAILED DESCRIPTION

Publications and patents are referred to throughout this disclosure. AllU.S. Patents cited herein are hereby incorporated by reference. Allpercentages, ratios, and proportions used herein are percent by weightunless otherwise specified.

Novel 6-aminoisoquinoline compounds and methods of using those compoundsto treat disease are provided.

“Alkyl” refers to a saturated aliphatic hydrocarbon including straightchain and branched chain groups. “Alkyl” may be exemplified by groupssuch as methyl, ethyl, n-propyl, isopropyl, n-butyl and the like. Alkylgroups may be substituted or unsubstituted. Substituents may also bethemselves substituted. When substituted, the substituent group ispreferably but not limited to C₁-C₄ alkyl, aryl, amino, cyano, halogen,alkoxy or hydroxyl. “C₁-C₄ alkyl” refers to alkyl groups containing oneto four carbon atoms.

“Alkenyl” refers to an unsaturated aliphatic hydrocarbon moietyincluding straight chain and branched chain groups. Alkenyl moietiesmust contain at least one alkene. “Alkenyl” may be exemplified by groupssuch as ethenyl, n-propenyl, isopropenyl, n-butenyl and the like.Alkenyl groups may be substituted or unsubstituted. Substituents mayalso be themselves substituted. When substituted, the substituent groupis preferably alkyl, halogen or alkoxy. Substitutients may also bethemselves substituted. Substituents be placed on the alkene itself andalso on the adjacent member atoms or the alkynyl moiety “C₂-C₄ alkenyl”refers to alkenyl groups containing two to four carbon atoms.

“Alkynyl” refers to an unsaturated aliphatic hydrocarbon moietyincluding straight chain and branched chain groups. Alkynyl moietiesmust contain at least one alkyne. “Alkynyl” may be exemplified by groupssuch as ethynyl, propynyl, n-butynyl and the like. Alkynyl groups may besubstituted or unsubstituted. When substituted, the substituent group ispreferably alkyl, amino, cyano, halogen, alkoxyl or hydroxyl.Substituents may also be themselves substituted. Substituents are not onthe alkyne itself but on the adjacent member atoms of the alkynylmoiety. “C₂-C₄ alkynyl” refers to alkynyl groups containing two to fourcarbon atoms.

“Acyl” or “carbonyl” refers to the group —C(O)R wherein R is alkyl;alkenyl; alkyl alkynyl, aryl, heteroaryl, carbocyclic,heterocarbocyclic; C₁-C₄ alkyl aryl or C₁-C₄ alkyl heteroaryl. C₁-C₄alkylcarbonyl refers to a group wherein the carbonyl moiety is precededby an alkyl chain of 1-4 carbon atoms.

“Alkoxy” refers to the group —O—R wherein R is acyl, alkyl alkenyl,alkyl alkynyl, aryl, carbocyclic; heterocarbocyclic; heteroaryl, C₁-C₄alkyl aryl or C₁-C₄ alkyl heteroaryl

“Amino” refers to the group —NR′R′ wherein each R′ is, independently,hydrogen, alkyl, aryl, heteroaryl, C₁-C₄ alkyl aryl or C₁-C₄ alkylheteroaryl. The two R′ groups may themselves be linked to form a ring.

“Aryl” refers to an aromatic carbocyclic group. “Aryl” may beexemplified by phenyl. The aryl group may be substituted orunsubstituted. Substituents may also be themselves substituted. Whensubstituted, the substituent group is preferably but not limited toheteroaryl; acyl, carboxyl, carbonylamino, nitro, amino, cyano, halogen,or hydroxyl.

“Carboxyl” refers to the group —C(═O)O—C₁-C₄ alkyl.

“Carbonylamino” refers to the group —C(O)NR′R′ wherein each R′ is,independently, hydrogen, alkyl, aryl, cycloalkyl; heterocycloalkyl;heteroaryl, C₁-C₄ alkyl aryl or C₁-C₄ alkyl heteroaryl. The two R′groups may themselves be linked to form a ring.

“C₁-C₄ alkyl aryl” refers to C₁-C₄ alkyl groups having an arylsubstituent such that the aryl substituent is bonded through an alkylgroup. “C₁-C₄ alkyl aryl” may be exemplified by benzyl.

“C₁-C₄ alkyl heteroaryl” refers to C₁-C₄ alkyl groups having aheteroaryl substituent such that the heteroaryl substituent is bondedthrough an alkyl group.

“Carbocyclic group” or “cycloalkyl” means a monovalent saturated orunsaturated hydrocarbon ring. Carbocyclic groups are monocyclic, or arefused, spiro, or bridged bicyclic ring systems. Monocyclic carbocyclicgroups contain 3 to 10 carbon atoms, preferably 4 to 7 carbon atoms, andmore preferably 5 to 6 carbon atoms in the ring. Bicyclic carbocyclicgroups contain 8 to 12 carbon atoms, preferably 9 to 10 carbon atoms inthe ring. Carbocyclic groups may be substituted or unsubstituted.Substituents may also be themselves substituted. Preferred carbocyclicgroups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cyclohexenyl, and cycloheptyl. More preferred carbocyclic groups includecyclopropyl and cyclobutyl. The most preferred carbocyclic group iscyclopropyl. Carbocyclic groups are not aromatic.

“Halogen” refers to fluoro, chloro, bromo or iodo moieties. Preferably,the halogen is fluoro, chloro, or bromo.

“Heteroaryl” or “heteroaromatic” refers to a monocyclic or bicyclicaromatic carbocyclic radical having one or more heteroatoms in thecarbocyclic ring. Heteroaryl may be substituted or unsubstituted. Whensubstituted, the substituents may themselves be substituted. Preferredbut non limiting substituents are aryl; C₁-C₄ alkylaryl; amino; halogen,hydroxy, cyano, nitro; carboxyl; carbonylamino or C₁-C₄ alkyl. Preferredheteroaromatic groups include tetrazoyl, triazolyl; thienyl, thiazolyl,purinyl, pyrimidyl, pyridyl, and furanyl. More preferred heteroaromaticgroups include benzothiofuranyl; thienyl, furanyl, tetrazoyl, triazolyl;and pyridyl.

“Heteroatom” means an atom other than carbon in the ring of aheterocyclic group or a heteroaromatic group or the chain of aheterogeneous group. Preferably, heteroatoms are selected from the groupconsisting of nitrogen, sulfur, and oxygen atoms. Groups containing morethan one heteroatom may contain different heteroatoms.

“Heterocarbocyclic group” or “heterocycloalkyl” or “heterocyclic” meansa monovalent saturated or unsaturated hydrocarbon ring containing atleast one heteroatom. Heterocarbocyclic groups are monocyclic, or arefused, spiro, or bridged bicyclic ring systems. Monocyclicheterocarbocyclic groups contain 3 to 10 carbon atoms, preferably 4 to 7carbon atoms, and more preferably 5 to 6 carbon atoms in the ring.Bicyclic heterocarbocyclic groups contain 8 to 12 carbon atoms,preferably 9 to 10 carbon atoms in the ring. Heterocarbocyclic groupsmay be substituted or unsubstituted. Substituents may also be themselvessubstituted. Preferred heterocarbocyclic groups include epoxy,tetrahydrofuranyl, azacyclopentyl, azacyclohexyl, piperidyl, andhomopiperidyl. More preferred heterocarbocyclic groups includepiperidyl, and homopiperidyl. The most preferred heterocarbocyclic groupis piperidyl. Heterocarbocyclic groups are not aromatic.

“Hydroxy” or “hydroxyl” means a chemical entity that consists of —OH.Alcohols contain hydroxy groups. Hydroxy groups may be free orprotected. An alternative name for hydroxyl is hydroxy

“Linker” means a linear chain of n member atoms where n is an integer offrom 1 to 4.

“Member atom” means a carbon, nitrogen, oxygen or sulfur atom. Memberatoms may be substituted up to their normal valence. If substitution isnot specified the substituents required for valency are hydrogen.

“Ring” means a collection of member atoms that are cyclic. Rings may becarbocyclic, aromatic, or heterocyclic or heteroaromatic, and may besubstituted or unsubstituted, and may be saturated or unsaturated. Ringjunctions with the main chain may be fused or spirocyclic. Rings may bemonocyclic or bicyclic. Rings contain at least 3 member atoms and atmost 10 member atoms. Monocyclic rings may contain 3 to 7 member atomsand bicyclic rings may contain from 8 to 12 member atoms. Bicyclic ringsthemselves may be fused or spirocyclic.

“Thioalkyl” refers to the group —S-alkyl.

“Sulfonyl” refers to the —S(O)₂R′ group wherein R′ is alkoxy, alkyl,aryl, carbocyclic, heterocarbocyclic; heteroaryl, C₁-C₄ alkyl aryl orC₁-C₄ alkyl heteroaryl.

“Sulfonylamino” refers to the —S(O)₂NR′R′ group wherein each R′ isindependently alkyl, aryl, heteroaryl, C₁-C₄ alkyl aryl or C₁-C₄ alkylheteroaryl.

“Pharmaceutically acceptable carrier” means a carrier that is useful forthe preparation of a pharmaceutical composition that is: generallycompatible with the other ingredients of the composition, notdeleterious to the recipient, and neither biologically nor otherwiseundesirable. “A pharmaceutically acceptable carrier” includes both oneand more than one carrier. Embodiments include carriers for topical,ocular, parenteral, intravenous, intraperitoneal intramuscular,sublingual, nasal and oral administration. “Pharmaceutically acceptablecarrier” also includes agents for preparation of aqueous dispersions andsterile powders for injection or dispersions.

“Excipient” as used herein includes physiologically compatible additivesuseful in preparation of a pharmaceutical composition. Examples ofpharmaceutically acceptable carriers and excipients can for example befound in Remington Pharmaceutical Science, 16^(th) Ed.

“Therapeutically effective amount” as used herein refers to a dosage ofthe compounds or compositions effective for influencing, reducing orinhibiting the activity of or preventing activation of a kinase. Thisterm as used herein may also refer to an amount effective at bringingabout a desired in vivo effect in an animal, preferably, a human, suchas reduction in intraocular pressure.

“Administering” as used herein refers to administration of the compoundsas needed to achieve the desired effect.

“Eye disease” as used herein includes, but is not limited to, glaucoma,allergy, cancers of the eye, neurodegenerative diseases of the eye, anddry eye.

The term “disease or condition associated with kinase activity” is usedto mean a disease or condition treatable, in whole or in part, byinhibition of one or more kinases.

The term “controlling the disease or condition” is used to mean changingthe activity of one or more kinases to affect the disease or condition.

The 6-aminoisoquinoline compounds may be represented by Formula I:

wherein A is a substituted or unsubstituted linker consisting of atleast two member atoms and at most 4 member atoms wherein the linker maybe mono- or disubstituted with halogen, cyano, amino, nitro, alkyl,alkenyl or alkynyl, or the substituted atoms may attach back to the mainchain to form a ring, wherein R₃ is hydrogen; halogen; alkyl; alkenyl;alkynyl; alkoxy; amino; cyano; cycloalkyl; heterocycloalkyl; aryl; C₁-C₄alkyl aryl; heteroaryl; C₁-C₄ alkyl heteroaryl; carbonyl; carbonylamino;thioalkyl; sulfonyl; sulfonylamino; acyl; or carboxyl; and wherein R₁,and R₂ are, independently, hydrogen; hydroxyl, halogen; C₁-C₄ alkyl;C₂-C₄ alkenyl, C₂-C₄ alkynyl, amino; nitro; cyano; C₁-C₄ carbonyl; C₁-C₄carbonylamino; C₁-C₄ alkoxy; C₁-C₄ sulfonyl; C₁-C₄ sulfonylamino; C₁-C₄thioalkyl and C₁-C₄ carboxyl.

In a preferred embodiment of Formula (I), A is a cyclopropyl ring, R₃ isa substituted aromatic ring and R₁, R₂ are hydrogen. In anotherpreferred embodiment of Formula (I), A is —CH₂NH—. In another preferredembodiment of Formula (I), R₁, and R₂, are hydrogen and R₃ is asubstituted aromatic ring.

In another embodiment, the 6-aminoisoquinoline compounds may berepresented by Formula (II):

wherein R₁, and R₂ are, independently, hydrogen; hydroxyl, halogen;C₁-C₄ alkyl; C₂-C₄ alkenyl, C₂-C₄ alkynyl, amino; nitro; cyano; C₁-C₄carbonyl; C₁-C₄ carbonylamino; C₁-C₄ alkoxy; C₁-C₄ sulfonyl; C₁-C₄sulfonylamino; C₁-C₄ thioalkyl and C₁-C₄ carboxyl. andwherein R₄ is hydrogen; halogen; alkyl; alkenyl; alkynyl; alkoxy; amino;cyano; cycloalkyl; heterocycloalkyl; aryl; C₁-C₄ alkyl aryl; heteroaryl;C₁-C₄ alkyl heteroaryl; carbonyl; carbonylamino; thioalkyl; sulfonyl;sulfonylamino; acyl; or carboxyl; andB is a chain containing from 0 to 3 member atoms,X represents n independently chosen member atoms which together form aring structure and n is an integer from about 0 to about 5.

In some preferred embodiments of Formula II, the aminoisoquinolinesinclude those compounds wherein R₁ and R₂ are hydrogen and B contains 0(zero) member atoms. In further preferred embodiments, R₄ is a meta orpara-substituted aromatic ring. In some preferred embodiments R₄ is anaromatic ring that is substituted with a halogen in the ortho positionand a carbonyl group in the para position.

In another embodiment, the 6-aminoisoquinoline compounds may berepresented by Formula (III):

Wherein one X is independently and uniquely selected from CH₂, O, S,S(O), S(O)(O),

andThe other X groups are chosen from the same list so as to create astable moiety, or omitted from the formula,wherein R₁, and R₂ are, independently, hydrogen; hydroxyl, halogen;C₁-C₄ alkyl; C₂-C₄ alkenyl, C₂-C₄ alkynyl, amino; nitro; cyano; C₁-C₄carbonyl; C₁-C₄ carbonylamino; C₁-C₄ alkoxy; C₁-C₄ sulfonyl; C₁-C₄sulfonylamino; C₁-C₄ thioalkyl and C₁-C₄ carboxyl; andR₅ and R₁₀ are independently hydrogen; halogen; alkyl; alkenyl; alkynyl;alkoxy; amino; cyano; cycloalkyl; heterocycloalkyl; aryl; C₁-C₄ alkylaryl; heteroaryl; C₁-C₄ alkyl heteroaryl; carbonyl; carbonylamino;thioalkyl; sulfonyl; sulfonylamino; acyl; or carboxyl; and

A 6-aminoisoquinoline compound according to Formula IV is furtherprovided:

wherein A is a substituted or unsubstituted linker consisting of atleast one member atom and at most 4 member atoms wherein the linker maybe mono- or disubstituted with halogen, cyano, nitro or C₁-C₄ alkyl, orthe substituted atoms may attach back to the main chain to form a ring,andwherein R₃ is hydrogen; halogen; alkyl; alkenyl; alkynyl; alkoxy; amino;cyano; cycloalkyl; heterocycloalkyl; aryl; C₁-C₄ alkyl aryl; heteroaryl;C₁-C₄ alkyl heteroaryl; carbonyl; carbonylamino; thioalkyl; sulfonyl;sulfonylamino; acyl; or carboxyl; andwherein R₁, and R₂ are, independently, hydrogen; hydroxyl, halogen;C₁-C₄ alkyl; C₂-C₄ alkenyl, C₂-C₄ alkynyl, amino; nitro; cyano; C₁-C₄carbonyl; C₁-C₄ carbonylamino; C₁-C₄ alkoxy; C₁-C₄ sulfonyl; C₁-C₄sulfonylamino; C₁-C₄ thioalkyl and C₁-C₄ carboxyl.

In some preferred embodiments of Formula IV, the aminoisoquinolinesinclude those compounds wherein R₁ and R₂ are hydrogen and A is ethyl orethenyl. In further preferred embodiments, R₃ is a meta orpara-substituted aromatic ring. In some preferred embodiments R₃ is anaromatic ring that is substituted with a halogen in the ortho positionand a carbonyl group in the para position. In some preferred embodimentsA is —O—CH₂— or —NH—CH₂—.

The carbon linked 6-aminoisoquinoline compounds may be synthesized bythe general scheme set forth below:

Scheme One:

The selected aromatic ester (1) was saponified with an appropriate basesuch as LiOH to form the free acid (2) then coupled to6-aminoisoquinoline using standard coupling procedures such as EDC andDMAP in DMF to form the desired compound (3). Alternatively, the ester(1) was reacted with the trimethyl aluminum amide of 6-aminoisoquinolineto generate the amide (3) directly.

Intermediates (1) of the type III and IV from Scheme One may besynthesized by the general scheme, Scheme Two set forth below:

Scheme Two:

The selected ester is reacted with trimethylsulfoxonium ylide to givethe desired cyclopropyl intermediate (1) for use in Scheme One.

When intermediates (1) of the type III or IV are aromatic ringssubstituted in the meta or para positions with amides or esters, theyare synthesized by the general scheme set forth below:

Scheme Three:

The selected allylic ester was converted to a cyclopropane withtrimethyl sulfoxonium ylide. The aromatic acid group of the formedcyclopropane was then converted to its acid chloride using standardprocedures then reacted with the appropriate alcohol or amine to givethe desired intermediate (1).

Scheme Four:

The appropriate ester is converted to a cyclopropane under Simmons-Smithconditions to give the desired intermediate (1).

Using Scheme Five, compounds of Formula III with substituted 6-aminoisoquinolines (1) are prepared.

Scheme Five:

Chloroacetic acid is treated with EDC in the presence of DMAP and thenthe substituted 6-aminoisoquinoline. The resulting amide is treated withsodium iodide and an amine to provide the final product amine 4.

Using Scheme Six, the compound with a sulfonamide group according toFormula IV may be prepared.

Scheme Six:

6-Aminoisoquinoline is treated with sodium hydride and thenchloromethanesulfonyl chloride to provide the sulfonamide 5. Treatmentof the chlorosulfonamide 5 with an amine in the presence of sodiumiodide provides the desired amine 6.

Using reactions similar to those in Scheme Five, compounds with acycloalkyl moiety can be prepared. The compounds are obtained by using acycloalkylamine instead of an aniline to displace the chloride (SchemeSeven).

Scheme Seven:

Treatment of the chloroamide 7 with an amine in the presence of sodiumiodide provides the desired product 8.

The abbreviations used in the synthetic schemes shown have the followingmeanings: Boc₂O means di-tert-butyl-dicarbonate, DMAP means dimethylaminopyridine, DMSO means Dimethyl Sulfoxide, HATU means2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate, LDA means lithium diisopropyl amide, DMF isdimethylformamide, THF is tetrahydrofuran, and EDC meansN-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride.

The 6-aminoisoquinoline compounds of the above Formulae and compositionsincluding them have kinase inhibitory activity and are thus useful ininfluencing or inhibiting the action of kinases, and in treatment and/orprevention of diseases or conditions influenced by kinases. The6-aminoisoquinolines may be used to influence or inhibit the action ofkinases either in a cell in vitro or in a cell in a living body in vivo.Specifically, in one embodiment, a method is provided of inhibiting theaction of a kinase comprising applying to a medium such as an assaymedium or contacting with a cell either in a cell in vitro or in a cellin a living body in vivo an effective inhibitory amount of a compoundaccording to Formula (I) or (II) or (III) or (IV). In a preferredembodiment, the kinase inhibited is a rho kinase. Compounds according toFormula (I) or (II) or (III) or (IV) are used in methods of inhibitingkinases in a cell, a tissue or an animal such as a human comprisingadministering to, or contacting with, the cell a therapeuticallyeffective amount of one or more of these 6-aminoisoquinolines. The oneor more of the 6-aminoisoquinolines are preferably administered in apharmaceutically acceptable formulation, such as in or with apharmaceutically acceptable carrier when the 6-aminoisoquinolines areadministered to a cell or cells in a living organism or body. In anotherembodiment, the 6-aminoisoquinolines according to Formula (I) or (II) or(III) or (IV) are used in methods for influencing the action of a kinasein a cell comprising administering to, or contacting with, the cell aneffective amount of one or more 6-aminoisoquinolines for influencing theaction of the kinase in the cell. The one or more of the6-aminoisoquinolines are preferably administered in a pharmaceuticallyacceptable formulation, such as in or with a pharmaceutically acceptablecarrier when the 6-aminoisoquinolines are administered to a cell orcells in a living organism or body.

Treatment or prevention of diseases or conditions for which the6-aminoisoquinolines may be useful includes any of the diseases orconditions associated with kinase activity or diseases or conditionsaffected by kinases. Examples of these types of diseases include retinaldegradation, glaucoma and cancer.

The 6-aminoisoquinolines in some embodiments will be administered inconjunction with the administration of a second or in some cases a thirdtherapeutic agent which is directed to the treatment or prevention of acondition or disease affected by those specific receptors. Combiningadministration of the 6-aminoisoquinolines with other therapeutic agentswill provide a reduction or prevention of the condition or disease towhich the therapeutic agent is directed, resulting in improving theability of the therapeutic agent to have the desired effect over alonger period of time. Additionally, the administration of thetherapeutic agent or receptor agonist with an 6-aminoisoquinolineformulation will enable lower doses of the other therapeutic agents tobe administered for a longer period of time.

One or more therapeutic agents may be administered with one or more6-aminoisoquinoline compounds. The therapeutic agents and/or the6-aminoisoquinoline compounds are preferably administered in apharmaceutically acceptable formulation with a pharmaceuticallyacceptable carrier when the 6-aminoisoquinolines are administered to acell or cells in a living organism or a mammal, preferably human.

Compositions including the 6-aminoisoquinolines of Formula (I) or (II)or (III) or (IV) may be obtained in the form of various salts orsolvates. As the salts, physiologically acceptable salts or saltsavailable as raw materials are used.

Compositions may include one or more of the isoforms of Formula (I) or(II) or (III) or (IV) when present. When a stereocenter exists, eachenantiomer may be separately used, or they may be combined in anyproportion.

Pharmaceutical compositions for use in accordance with the presentinvention may be formulated in a conventional manner using one or morephysiologically acceptable carriers or excipients. Thus, the compoundsand their physiologically acceptable salts and solvates may beformulated for administration by, for example, solid dosing, eyedrop, ina topical oil-based formulation, injection, inhalation (either throughthe mouth or the nose), oral, buccal, parenteral or rectaladministration. Techniques and formulations may generally be found in“Reminington's Pharmaceutical Sciences”, (Meade Publishing Co., Easton,Pa.). Therapeutic compositions must typically be sterile and stableunder the conditions of manufacture and storage.

Compositions of the present invention may comprise a safe and effectiveamount of the subject compounds, and a pharmaceutically-acceptablecarrier. As used herein, “safe and effective amount” means an amount ofa compound sufficient to significantly induce a positive modification inthe condition to be treated, but low enough to avoid serious sideeffects (at a reasonable benefit/risk ratio), within the scope of soundmedical judgment. A safe and effective amount of a compound will varywith the particular condition being treated, the age and physicalcondition of the patient being treated, the severity of the condition,the duration of the treatment, the nature of concurrent therapy, theparticular pharmaceutically-acceptable carrier utilized, and likefactors within the knowledge and expertise of the attending physician.

The route by which the A compound of the present invention (component A)will be administered and the form of the composition will dictate thetype of carrier (component B) to be used. The composition may be in avariety of forms, suitable, for example, for systemic administration(e.g., oral, rectal, nasal, sublingual, buccal, or parenteral) ortopical administration (e.g., local application on the skin, ocular,liposome delivery systems, or iontophoresis).

Carriers for systemic administration typically comprise at least one ofa) diluents, b) lubricants, c) binders, d) disintegrants, e) colorants,f) flavors, g) sweeteners, h) antioxidants, j) preservatives, k)glidants, m) solvents, n) suspending agents, o) wetting agents, p)surfactants, combinations thereof, and others. All carriers are optionalin the systemic compositions.

Ingredient a) is a diluent. Suitable diluents for solid dosage formsinclude sugars such as glucose, lactose, dextrose, and sucrose; diolssuch as propylene glycol; calcium carbonate; sodium carbonate; sugaralcohols, such as glycerin; mannitol; and sorbitol. The amount ofingredient a) in the systemic or topical composition is typically about50 to about 90%.

Ingredient b) is a lubricant. Suitable lubricants for solid dosage formsare exemplified by solid lubricants including silica, talc, stearic acidand its magnesium salts and calcium salts, calcium sulfate; and liquidlubricants such as polyethylene glycol and vegetable oils such as peanutoil, cottonseed oil, sesame oil, olive oil, corn oil and oil oftheobroma. The amount of ingredient b) in the systemic or topicalcomposition is typically about 5 to about 10%.

Ingredient c) is a binder. Suitable binders for solid dosage formsinclude polyvinylpyrrolidone; magnesium aluminum silicate; starches suchas corn starch and potato starch; gelatin; tragacanth; and cellulose andits derivatives, such as sodium carboxymethylcellulose, ethyl cellulose,methylcellulose, microcrystalline cellulose, and sodiumcarboxymethylcellulose. The amount of ingredient c) in the systemiccomposition is typically about 5 to about 50%, and in ocular soliddosing forms up to 99%.

Ingredient d) is a disintegrant. Suitable disintegrants for solid dosageforms include agar, alginic acid and the sodium salt thereof,effervescent mixtures, croscarmelose, crospovidone, sodium carboxymethylstarch, sodium starch glycolate, clays, and ion exchange resins. Theamount of ingredient d) in the systemic or topical composition istypically about 0.1 to about 10%.

Ingredient e) for solid dosage forms is a colorant such as an FD&C dye.When used, the amount of ingredient e) in the systemic or topicalcomposition is typically about 0.005 to about 0.1%.

Ingredient f) for solid dosage forms is a flavor such as menthol,peppermint, and fruit flavors. The amount of ingredient f), when used,in the systemic or topical composition is typically about 0.1 to about1.0%.

Ingredient g) for solid dosage forms is a sweetener such as aspartameand saccharin. The amount of ingredient g) in the systemic or topicalcomposition is typically about 0.001 to about 1%.

Ingredient h) is an antioxidant such as butylated hydroxyanisole(“BHA”), butylated hydroxytoluene (“BHT”), and vitamin E. The amount ofingredient h) in the systemic or topical composition is typically about0.1 to about 5%.

Ingredient j) is a preservative such as benzalkonium chloride, methylparaben and sodium benzoate. The amount of ingredient j) in the systemicor topical composition is typically about 0.01 to about 5%.

Ingredient k) for solid dosage forms is a glidant such as silicondioxide. The amount of ingredient k) in the systemic or topicalcomposition is typically about 1 to about 5%.

Ingredient m) is a solvent, such as water, isotonic saline, ethyloleate, glycerine, hydroxylated castor oils, alcohols such as ethanol,and phosphate buffer solutions. The amount of ingredient m) in thesystemic or topical composition is typically from about 0 to about 100%.

Ingredient n) is a suspending agent. Suitable suspending agents includeAVICEL® RC-591 (from FMC Corporation of Philadelphia, Pa.) and sodiumalginate. The amount of ingredient n) in the systemic or topicalcomposition is typically about 1 to about 8%.

Ingredient o) is a surfactant such as lecithin, polysorbate 80, andsodium lauryl sulfate, and the TWEENS® from Atlas Powder Company ofWilmington, Del. Suitable surfactants include those disclosed in theC.T.F.A. Cosmetic Ingredient Handbook, 1992, pp. 587-592; Remington'sPharmaceutical Sciences, 15th Ed. 1975, pp. 335-337; and McCutcheon'sVolume 1, Emulsifiers & Detergents, 1994, North American Edition, pp.236-239. The amount of ingredient o) in the systemic or topicalcomposition is typically about 0.1% to about 5%.

Although the amounts of components A and B in the systemic compositionswill vary depending on the type of systemic composition prepared, thespecific derivative selected for component A and the ingredients ofcomponent B, in general, system compositions comprise 0.01% to 50% ofcomponent A and 50 to 99.99% of component B.

Compositions for parenteral administration typically comprise A) 0.1 to10% of the compounds of the present invention and B) 90 to 99.9% of acarrier comprising a) a diluent and m) a solvent. In one embodiment,component a) comprises propylene glycol and m) comprises ethanol orethyl oleate.

Compositions for oral administration can have various dosage forms. Forexample, solid forms include tablets, capsules, granules, and bulkpowders. These oral dosage forms comprise a safe and effective amount,usually at least about 5%, and more particularly from about 25% to about50% of component A). The oral dosage compositions further comprise about50 to about 95% of component B), and more particularly, from about 50 toabout 75%.

Tablets can be compressed, tablet triturates, enteric-coated,sugar-coated, film-coated, or multiple-compressed. Tablets typicallycomprise component A, and component B a carrier comprising ingredientsselected from the group consisting of a) diluents, b) lubricants, c)binders, d) disintegrants, e) colorants, f) flavors, g) sweeteners, k)glidants, and combinations thereof. Specific diluents include calciumcarbonate, sodium carbonate, mannitol, lactose and cellulose. Specificbinders include starch, gelatin, and sucrose. Specific disintegrantsinclude alginic acid and croscarmelose. Specific lubricants includemagnesium stearate, stearic acid, and talc. Specific colorants are theFD&C dyes, which can be added for appearance. Chewable tabletspreferably contain g) sweeteners such as aspartame and saccharin, or f)flavors such as menthol, peppermint, fruit flavors, or a combinationthereof.

Capsules (including time release and sustained release formulations)typically comprise component A, and a carrier comprising one or more a)diluents disclosed above in a capsule comprising gelatin. Granulestypically comprise component A, and preferably further comprise k)glidants such as silicon dioxide to improve flow characteristics.

The selection of ingredients in the carrier for oral compositionsdepends on secondary considerations like taste, cost, and shelfstability, which are not critical for the purposes of this invention.One skilled in the art would know how to select appropriate ingredientswithout undue experimentation.

The solid compositions may also be coated by conventional methods,typically with pH or time-dependent coatings, such that component A isreleased in the gastrointestinal tract in the vicinity of the desiredapplication, or at various points and times to extend the desiredaction. The coatings typically comprise one or more components selectedfrom the group consisting of cellulose acetate phthalate, polyvinylacetate phthalate, hydroxypropyl methyl cellulose phthalate, ethylcellulose, EUDRAGIT® coatings (available from Rohm & Haas G. M. B. H. ofDarmstadt, Germany), waxes and shellac.

Compositions for oral administration can also have liquid forms. Forexample, suitable liquid forms include aqueous solutions, emulsions,suspensions, solutions reconstituted from non-effervescent granules,suspensions reconstituted from non-effervescent granules, effervescentpreparations reconstituted from effervescent granules, elixirs,tinctures, syrups, and the like. Liquid orally administered compositionstypically comprise component A and component B, namely, a carriercomprising ingredients selected from the group consisting of a)diluents, e) colorants, f) flavors, g) sweeteners, j) preservatives, m)solvents, n) suspending agents, and o) surfactants. Peroral liquidcompositions preferably comprise one or more ingredients selected fromthe group consisting of e) colorants, f) flavors, and g) sweeteners.

Other compositions useful for attaining systemic delivery of the subjectcompounds include sublingual, buccal and nasal dosage forms. Suchcompositions typically comprise one or more of soluble filler substancessuch as a) diluents including sucrose, sorbitol and mannitol; and c)binders such as acacia, microcrystalline cellulose, carboxymethylcellulose, and hydroxypropyl methylcellulose. Such compositions mayfurther comprise b) lubricants, e) colorants, f) flavors, g) sweeteners,h) antioxidants, and k) glidants.

In one embodiment of the invention, the compounds of the presentinvention are topically administered. Topical compositions that can beapplied locally to the eye may be in any form known in the art,non-limiting examples of which include solids, gelable drops, sprays,ointments, or a sustained or non-sustained release unit placed in theconjunctival cul-du-sac of the eye or another appropriate location.

Topical compositions that can be applied locally to the skin may be inany form including solids, solutions, oils, creams, ointments, gels,lotions, shampoos, leave-on and rinse-out hair conditioners, milks,cleansers, moisturizers, sprays, skin patches, and the like. Topicalcompositions comprise: component A, the compounds described above, andcomponent B, a carrier. The carrier of the topical compositionpreferably aids penetration of the compounds into the eye. Component Bmay further comprise one or more optional components.

The dosage range of the compound for systemic administration is fromabout 0.01 to about 1000 μg/kg body weight, preferably from about 0.1 toabout 100 μg/kg per body weight, most preferably form about 1 to about50 μg/kg body weight per day. The transdermal dosages will be designedto attain similar serum or plasma levels, based upon techniques known tothose skilled in the art of pharmacokinetics and transdermalformulations. Plasma levels for systemic administration are expected tobe in the range of 0.01 to 100 nanograms/mL, (ng/mL) more preferablyfrom 0.05 to 50 ng/mL and most preferably from 0.1 to 10 ng/mL. Whilethese dosages are based upon a daily administration rate, weekly ormonthly accumulated dosages may also be used to calculate the clinicalrequirements.

Dosages may be varied based on the patient being treated, the conditionbeing treated, the severity of the condition being treated, the route ofadministration, etc. to achieve the desired effect.

The compounds of the present invention are useful in decreasingintraocular pressure. Thus, these compounds are useful in the treatmentof glaucoma. The preferred route of administration for treating glaucomais topically.

The exact amounts of each component in the topical composition depend onvarious factors. The amount of component A added to the topicalcomposition is dependent on the IC₅₀ of component A, typically expressedin nanomolar (nM) units. For example, if the IC₅₀ of the medicament is 1nM, the amount of component A will be from about 0.0001 to about 0.1%.If the IC₅₀ of the medicament is 10 nM, the amount of component A) willbe from about 0.01 to about 1%. If the IC₅₀ of the medicament is 100 nM,the amount of component A will be from about 0.1 to about 10%. If theIC₅₀ of the medicament is 1000 nM, the amount of component A will be 1to 100%, preferably 5% to 50%. If the amount of component A is outsidethe ranges specified above (i.e., lower), efficacy of the treatment maybe reduced. One skilled in the art would know how to calculate an IC₅₀.The remainder of the composition, up to 100%, is component B.

The amount of the carrier employed in conjunction with component A issufficient to provide a practical quantity of composition foradministration per unit dose of the medicament. Techniques andcompositions for making dosage forms useful in the methods of thisinvention are described in the following references: ModernPharmaceutics, Chapters 9 and 10, Banker & Rhodes, eds. (1979);Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1981); andAnsel, Introduction to Pharmaceutical Dosage Forms, 2^(nd) Ed., (1976).

Component B may comprise a single ingredient or a combination of two ormore ingredients. In the topical compositions, component B comprises atopical carrier. Suitable topical carriers comprise one or moreingredients selected from the group consisting of phosphate bufferedsaline, isotonic water, deionized water, monofunctional alcohols,symmetrical alcohols, aloe vera gel, allantoin, glycerin, vitamin A andE oils, mineral oil, propylene glycol, PPG-2 myristyl propionate,dimethyl isosorbide, castor oil, combinations thereof, and the like.More particularly, carriers for skin applications include propyleneglycol, dimethyl isosorbide, and water, and even more particularly,phosphate buffered saline, isotonic water, deionized water,monofunctional alcohols and symmetrical alcohols.

The carrier of the topical composition may further comprise one or moreingredients selected from the group consisting of q) emollients, r)propellants, s) solvents, t) humectants, u) thickeners, v) powders, w)fragrances, x) pigments, and y) preservatives.

Ingredient q) is an emollient. The amount of ingredient q) in askin-based topical composition is typically about 5 to about 95%.Suitable emollients include stearyl alcohol, glyceryl monoricinoleate,glyceryl monostearate, propane-1,2-diol, butane-1,3-diol, mink oil,cetyl alcohol, isopropyl isostearate, stearic acid, isobutyl palmitate,isocetyl stearate, oleyl alcohol, isopropyl laurate, hexyl laurate,decyl oleate, octadecan-2-ol, isocetyl alcohol, cetyl palmitate,di-n-butyl sebacate, isopropyl myristate, isopropyl palmitate, isopropylstearate, butyl stearate, polyethylene glycol, triethylene glycol,lanolin, sesame oil, coconut oil, arachis oil, castor oil, acetylatedlanolin alcohols, petroleum, mineral oil, butyl myristate, isostearicacid, palmitic acid, isopropyl linoleate, lauryl lactate, myristyllactate, decyl oleate, myristyl myristate, and combinations thereof.Specific emollients for skin include stearyl alcohol andpolydimethylsiloxane.

Ingredient r) is a propellant. The amount of ingredient r) in thetopical composition is typically about 0 to about 95%. Suitablepropellants include propane, butane, isobutane, dimethyl ether, carbondioxide, nitrous oxide, and combinations thereof.

Ingredient s) is a solvent. The amount of ingredient s) in the topicalcomposition is typically about 0 to about 95%. Suitable solvents includewater, ethyl alcohol, methylene chloride, isopropanol, castor oil,ethylene glycol monoethyl ether, diethylene glycol monobutyl ether,diethylene glycol monoethyl ether, dimethylsulfoxide, dimethylformamide, tetrahydrofuran, and combinations thereof. Specific solventsinclude ethyl alcohol and homotopic alcohols.

Ingredient t) is a humectant. The amount of ingredient t) in the topicalcomposition is typically 0 to 95%. Suitable humectants include glycerin,sorbitol, sodium 2-pyrrolidone-5-carboxylate, soluble collagen, dibutylphthalate, gelatin, and combinations thereof. Specific humectantsinclude glycerin.

Ingredient u) is a thickener. The amount of ingredient u) in the topicalcomposition is typically about 0 to about 95%.

Ingredient v) is a powder. The amount of ingredient v) in the topicalcomposition is typically 0 to 95%. Suitable powders includebeta-cyclodextrins, hydroxypropyl cyclodextrins, chalk, talc, fullersearth, kaolin, starch, gums, colloidal silicon dioxide, sodiumpolyacrylate, tetra alkyl ammonium smectites, trialkyl aryl ammoniumsmectites, chemically-modified magnesium aluminum silicate,organically-modified montmorillonite clay, hydrated aluminum silicate,fumed silica, carboxyvinyl polymer, sodium carboxymethyl cellulose,ethylene glycol monostearate, and combinations thereof. For ocularapplications, specific powders include beta-cyclodextrin, hydroxypropylcyclodextrin, and sodium polyacrylate. For gel dosing ocularformulations, sodium polyacrylate may be used.

Ingredient w) is a fragrance. The amount of ingredient w) in the topicalcomposition is typically about 0 to about 0.5%, particularly, about0.001 to about 0.1%. For ocular applications a fragrance is nottypically used.

Ingredient x) is a pigment. Suitable pigments for skin applicationsinclude inorganic pigments, organic lake pigments, pearlescent pigments,and mixtures thereof. Inorganic pigments useful in this inventioninclude those selected from the group consisting of rutile or anatasetitanium dioxide, coded in the Color Index under the reference CI77,891; black, yellow, red and brown iron oxides, coded under referencesCI 77,499, 77,492 and, 77,491; manganese violet (CI 77,742); ultramarineblue (CI 77,007); chromium oxide (CI 77,288); chromium hydrate (CI77,289); and ferric blue (CI 77,510) and mixtures thereof.

The organic pigments and lakes useful in this invention include thoseselected from the group consisting of D&C Red No. 19 (CI 45,170), D&CRed No. 9 (CI 15,585), D&C Red No. 21 (CI 45,380), D&C Orange No. 4 (CI15,510), D&C Orange No. 5 (CI 45,370), D&C Red No. 27 (CI 45,410), D&CRed No. 13 (CI 15,630), D&C Red No. 7 (CI 15,850), D&C Red No. 6 (CI15,850), D&C Yellow No. 5 (CI 19,140), D&C Red No. 36 (CI 12,085), D&COrange No. 10 (CI 45,425), D&C Yellow No. 6 (CI 15,985), D&C Red No. 30(CI 73,360), D&C Red No. 3 (CI 45,430), the dye or lakes based onCochineal Carmine (CI 75,570) and mixtures thereof.

The pearlescent pigments useful in this invention include those selectedfrom the group consisting of the white pearlescent pigments such as micacoated with titanium oxide, bismuth oxychloride, colored pearlescentpigments such as titanium mica with iron oxides, titanium mica withferric blue, chromium oxide and the like, titanium mica with an organicpigment of the above-mentioned type as well as those based on bismuthoxychloride and mixtures thereof. The amount of pigment in the topicalcomposition is typically about 0 to about 10%. For ocular applications apigment is generally not used.

In a particularly preferred embodiment of the invention, topicalpharmaceutical compositions for ocular administration are preparedtypically comprising component A and B (a carrier), such as purifiedwater, and one or more ingredients selected from the group consisting ofy) sugars or sugar alcohols such as dextrans, particularly mannitol anddextran 70, z) cellulose or a derivative thereof, aa) a salt, bb)disodium EDTA (Edetate disodium), and cc) a pH adjusting additive.

Examples of z) cellulose derivatives suitable for use in the topicalpharmaceutical composition for ocular administration include sodiumcarboxymethylcellulose, ethylcellulose, methylcellulose, andhydroxypropyl-methylcellulose, particularly,hydroxypropyl-methylcellulose.

Examples of aa) salts suitable for use in the topical pharmaceuticalcomposition for ocular administration include mono-, di- and trisodiumphosphate, sodium chloride, potassium chloride, and combinationsthereof.

Examples of cc) pH adjusting additives include HCl or NaOH in amountssufficient to adjust the pH of the topical pharmaceutical compositionfor ocular administration to 5.8-7.5.

Component A may be included in kits comprising component A, a systemicor topical composition described above, or both; and information,instructions, or both that use of the kit will provide treatment forcosmetic and medical conditions in mammals (particularly humans). Theinformation and instructions may be in the form of words, pictures, orboth, and the like. In addition or in the alternative, the kit maycomprise the medicament, a composition, or both; and information,instructions, or both, regarding methods of application of medicament,or of composition, preferably with the benefit of treating or preventingcosmetic and medical conditions in mammals (e.g., humans).

The invention will be further explained by the following illustrativeexamples that are intended to be non-limiting.

Procedures for preparation of the 6-aminoisoquinolines are described inthe following examples.

All temperatures are given in degrees Centigrade. Reagents and startingmaterials were purchased from commercial sources or prepared followingpublished literature procedures.

Unless otherwise noted, HPLC purification, when appropriate, wasperformed by redissolving the compound in a small volume of DMSO andfiltering through a 0.45 micron (nylon disc) syringe filter. Thesolution was then purified using, for example, a 50 mm Varian DynamaxHPLC 21.4 mm Microsorb Guard-8 C₈ column. A typical initial elutingmixture of 40-80% MeOH:H₂O was selected as appropriate for the targetcompound. This initial gradient was maintained for 0.5 minutes thenincreased to 100% MeOH:0% H₂O over 5 minutes. 100% MeOH was maintainedfor 2 more minutes before re-equilibration back to the initial startinggradient. A typical total run time was 8 minutes. The resultingfractions were analyzed, combined as appropriate, and then evaporated toprovide purified material.

Proton magnetic resonance (¹H NMR) spectra were recorded on either aVarian INOVA 400 MHz (¹H) NMR spectrometer, Varian INOVA 500 MHz (¹H)NMR spectrometer, Bruker ARX 300 MHz (¹H) NMR spectrometer, Bruker DPX400 MHz (¹H) NMR spectrometer, or a Bruker DRX 500 MHz (¹H) NMRspectrometer. All spectra were determined in the solvents indicated.Although chemical shifts are reported in ppm downfield oftetramethylsilane, they are referenced to the residual proton peak ofthe respective solvent peak for ¹H NMR. Interproton coupling constantsare reported in Hertz (Hz). Analytical HPLC was performed using aPhenomenex Aqua 5 micron C₁₈ 125 Å 50×4.60 mm column coupled with anAgilent 1100 series VWD UV detector. A neutral 0.1% BES (w/v) pH 7.1buffer with LiOH and 1% CH₃CN in H₂O is used as the aqueous phase. Theinitial gradient was 55% MeOH aqueous buffer which was increased to 100%MeOH over 3 minutes. 100% MeOH was maintained for 2 minutes before itwas re-equilibrated to the initial starting gradient. Spectra wereanalyzed at 254 nm. LCMS spectra were obtained using a ThermofinniganAQA MS ESI instrument. The samples were passed through a Phenomenex Aqua5 micron C₁₈ 125 Å 50×4.60 mm column. The initial gradient was 55% MeOH:1% CH₃CN in H₂O which was increased to 100% MeOH over 3 minutes. 100%MeOH was maintained for 2 minutes before it was re-equilibrated to theinitial starting gradient. The spray setting for the MS probe was at 350μL/min with a cone voltage at 25 mV and a probe temperature at 450° C.

The following preparations illustrate procedures for the preparation ofintermediates and methods for the preparation of 6-aminoisoquinolines.

Example 1. Preparation of 2-chloro-N-(isoquinolin-6-yl) acetamide (E1)

To chloroacetic acid in DMF was added EDC, DMAP and 6-aminoisoquinoline.This mixture was stirred for 4 hours. The reaction was washed withNaHCO₃(sat), extracted with EtOAc, dried (Na₂SO₄), filtered andevaporated. Column chromatography (SiO₂, 5% MeOH/CH₂Cl₂) gave2-chloro-N-(isoquinolin-6-yl)acetamide (E1).

Example 2. Preparation of N-(isoquinolin-6-yl)-3-phenylpropanamide (E2)

To 6-aminoisoquinoline in DMF cooled to 0° C. was added NaH and thesolution was stirred for 30 minutes at 0° C. Then hydrocinnamoylchloride was added and the mixture was stirred for 4 hours at roomtemperature. The mixture was diluted with EtOAc, extracted withNaHCO₃(sat), dried (Na₂SO₄), filtered and evaporated. Columnchromatography (SiO₂, 10% hexanes/EtOAc) gaveN-(isoquilin-6-yl)-3-phenylpropanamide (E2).

Using largely the procedures set forth in Examples 1-2 and substitutingthe appropriate starting materials, the compounds 3-7 were made.

Example No. R 3 CH₃ 4 m-CH₂—C₆H₄—OMe 5

6

7 trans-CH═CH—C₆H₅Similarly, using largely the procedures set forth in Examples 1-2 andsubstituting the appropriate starting materials, the compounds 8-38 canbe made.

Example No. R 8, 9, 10, CH₂CH₃, CH₂CH₂CH₃, CH₂CH═CH₂, 11, 12 CH(CH₃)₂,C(CH₃)₃ 13, 14, 15 CH₂F, CHF₂, CF₃ 16, 17, 18, CH₂OH, CH₂OMe, CH₂OEt,CH₂OBn, 19, 20, 21 CH₂OAc, CH₂OBz 22, 23, 24, 25, 26, 27, 28, 29, 30

Example No. B X 31 CH₂ m-C₆H₄—Cl 32 CH₂ p-C₆H₄—F 33 CH₂—CH₂ m-C₆H₄—CO₂Me34 CH₂—CH₂ m-C₆H₄—CONH₂ 35 CH₂—CH₂ m-C₆H₄—CONHMe 36 CH═CH m-C₆H₄—CO₂Me37 CH₂CH₂ m-C₆H₄—CONHBn 38 CH═CH o-C₆H₄—Cl

Example 39. Preparation of tert-butyl2-(isoquinolin-6-ylamino)-2-oxoethyl carbamate (E39)

To N-Boc-glycine in DMF was added EDC, HOBT and 6-aminoisoquinoline.This mixture was stirred for 8 hours. The reaction was washed withNaHCO₃(sat), extracted with EtOAc, dried (Na₂SO₄), filtered andevaporated. Column chromatography (SiO₂, MeOH/CH₂Cl₂) gave tert-butyl2-(isoquinolin-6-ylamino)-2-oxoethyl carbamate (E39).

Example 40. Preparation of 2-amino-N-isoquinoline-6-yl-acetamidehydrochloride (E40)

To tert-butyl 2-(isoquinolin-6-ylamino)-2-oxoethyl carbamate (E39) inCH₂Cl₂ was added HCl (4M) in dioxane and the solution was stirredovernight at room temperature. The reaction was concentrated to give2-amino-N-isoquinoline-6-yl-acetamide dihydrochloride (E40).

Using largely the procedures set forth in Examples 39-40 andsubstituting the appropriate starting materials, the compounds 41-57 canbe made.

Example R¹—R² (together No. R¹ R² form a ring) 41 H Me — 42 H CH₂C₆H₅ —43 H p-CH₂C₆H₄OH — 44 H p-CH₂C₆H₄OMe — 45 H CH₂CO₂Me — 46 H CH₂CH₂CO₂Me— 47 H CH₂CONH₂ — 48 H CH₂CH₂CONH2 — 49 Me Me — 50, 51, 52 — —

Example No. X 53 OH 54 OMe 55 NHMe 56 NHC₆H5 57 m-NHC₆H₄—OMe

Example 58. Preparation of 2-(benzylamino)-N-(isoquinolin-6-yl)acetamide(E58)

To 2-chloro-N-(isoquinolin-6-yl)acetamide (E1) in MeOH was added KI andthe solution was heated to 60° C. for 40 minutes. The mixture was cooledto 45° C. and benzylamine was added and stirred at 45° C. After 2-4hours, the solvents were evaporated and the residue was taken up inEtOAc and extracted with NaHCO₃(sat). The organics were dried (Na₂SO₄),filtered and evaporated. Flash chromatography (SiO₂, 2% NH₃(2M) inMeOH/3% MeOH/CH₂Cl₂) gave purified2-(benzylamino)-N-(isoquinolin-6-yl)acetamide (E58).

Example 59. Example ofN-(isoquinolin-6-yl)-2-(3-methoxybenzylamino)acetamide (E59)

To m-chlorobenzaldehyde in MeOH was added2-amino-N-(isoquinolin-6-yl)acetamide dihydrochloride (from Example 40)and the pH was adjusted to 5 with DIPEA. The mixture was stirred at roomtemperature for 2 hours, then NaCNBH₃ was added and the pH was adjustedto −5.0 with acetic acid. The mixture was stirred 12 hours, quenchedwith Na₂CO₃(sat), extracted with EtOAc, dried (Na₂SO₄), filtered andevaporated to give N-(isoquinolin-6-yl)-2-(3-chlorobenzylamino)acetamide(E59).

Using largely the procedures set forth in Examples 58-59 andsubstituting the appropriate starting materials, the compounds 60-63were made.

Example No. n1 n2 R¹ 60 0 0 p-C₆H₄—CO₂Me 61 0 0 m-C₆H₄—F 62 0 0m-C₆H₄—OAc 63 0 0 m-C₆H₄—OCF₃

Similarly, using largely the procedures set forth in Examples 58-59 andsubstituting the appropriate starting materials, the compounds 64-86 canbe made.

Example No. n1 n2 R¹ 64 0 0 p-C₆H₄—OMe 65 0 0 o-C₆H₄—Cl 66 0 0m-C₆H₄—CONHMe 67 0 0

68 0 0 CH₃ 69 0 0 m-C₆H₄—CF₃ 70 0 0 p-C₆H₄—F 71 0 0 1,2,4-C₆H₂—Cl₃ 72 00 1,4-C₆H₃—Cl₂ 73 0 0 2,4-C₆H₃—Cl₂ 74 0 0

75 0 0

76 0 0

77 0 0

78 0 0

79 1 0 o,p-C₆H₃—Cl₂ 80 1 0 m-C₆H₄—CO₂Me 81 1 0 o-C₆H₄—F 82 1 0 C₆H₅ 83 20 C₆H₅ 84 0 1 p-C₆H₄—Cl 85 0 1 p-C₆H₄CONHC₆H₅ 86 0 1 o-C₆H₅

Example 87. Preparation of benzylisoquinolin-6-ylcarbamate (E87)

To 6-aminoisoquinoline in DMF at −40° C. was added NaH and solution waswarmed to 0° C. for 30 minutes. Then benzylchloroformate was added andthe reaction stirred at 0° C. for 2 hours. The solution was quenchedwith AcOH, poured into NaHCO₃(sat) and extracted with EtOAc, dried(Na₂SO₄), filtered and evaporated. Flash chromatography (SiO₂ 90%EtOAc/Hex) gave benzylisoquinolin-6-ylcarbamate. (E87).

Example 88. Preparation of 1-benzyl-3-(isoquinolin-6-yl)urea (E88)

To 6-aminoisoquinoline in DMF is added DMAP and benzyl isocyanate andthe solution was stirred at room temperature for 4 hours. The mixturewas poured into NaHCO₃ (sat), extracted with EtOAc, dried (Na₂SO₄),filtered and evaporated. Flash chromatography (SiO₂, 5% MeOH/CH₂Cl₂)provided 1-benzyl-3-(isoquinolin-6-yl) urea (E88).

Using largely the procedures set forth in Examples 87-88 andsubstituting the appropriate starting materials, the compounds 89-98were made.

Example No. n X R¹ 89 0 O p-C₆H₄—OMe 90 0 NH o-C₆H₄—CH₂C₆H₅ 91 0 NHm-C₆H₄—CH₂C₆H₅ 92 0 NH p-C₆H₄—CH₂C₆H₅ 93 0 NH C₆H₅ 94 0 NH p-C₆H₄—Cl 951 O o-C₆H₄—Cl 96 1 NH o-C₆H₄—Cl 97 1 NH

98 2 O —OCH₂CH₂C₆H₅

Similarly, using largely the procedures set forth in Examples 87-88 andsubstituting the appropriate starting materials, the compounds 99-104can be made.

Example No. n X R1  99 0 O C₆H₅ 100 0 O m-C₆H₄—Cl 101 0 O m-C₆H₄—F 102 0NH

103 1 NH m-C₆H₄—CONHMe 104 1 NH p-C₆H₄—Cl

Example 105. Preparation of N-(isoquinolin-6-yl)-2-morpholinoacetamide.(E105)

To 2-chloro-N-(isoquinolin-6-yl)acetamide (E1) in MeOH is added KI andthe solution is heated to 60° C. for 40 minutes. The mixture is cooledto 45° C. and morpholine is added and stirred at 45° C. After 2-4 hours,the solvents are evaporated and the residue is taken up in EtOAc andextracted with NaHCO_(3(sat)). The organics are dried (Na₂SO₄), filteredand evaporated. Flash chromatography (2% NH₃(2M)/MeOH/3% MeOH/CH₂Cl₂)gives N-(isoquinolin-6-yl)-2-morpholino acetamide (E105).

Using largely the procedure set forth in Example 105 and substitutingthe appropriate starting materials, the compounds 106-116 can be made.

Example No. X 106

107

108

109

110

111

113

114

115

116

Example 117. Synthesis of 4-(2-(isoquinolin-6-ylcarbamoyl)cyclopropyl)-N-(4-phenoxyphenyl)benzamide: (E117d)

A stirred suspension of 4-formylcinnamic acid (58.3 mmol) in 130 mLmethanol was protected from the atmosphere with a CaCl₂ guard tubeattached to the end of a condenser. The reaction mixture was maintainedbetween −50 and −45° C. [dry ice/acetone] while thionyl chloride (3equiv.) was added over 1 h using syringe pump (addition rate: 0.25ml/min). During addition the reaction thickened but stirring continued.Following addition, the reaction was allowed to stir and come to roomtemperature overnight. Volatiles were removed under reduced pressure(Rotovap). Dichloromethane was added. The reaction was stirred and waterwas carefully added. The mixture was transferred to a separatory funnel.The aqueous layer was removed and discarded. The organic phase waswashed consecutively with brine, sat. NaHCO₃ and brine. The organicphase was dried over MgSO₄, filtered and concentrated. The solid wasrecrystallized from CH₂Cl₂/hexanes to provide the intermediate methylester.

The methyl ester (9.31 mmol) was dissolved in 300 ml acetone thendiluted with 75 ml water. To this suspension, the sulfamic acid (1.45eq.) and sodium chlorite (1.40 eq.) were added. The reaction stirred atroom temperature for three hours. The reaction mixture was extractedwith EtOAc and the combined organic extracts were dried over MgSO₄,filtered and concentrated. The material was recrystallized from EtOAc togive the intermediate acid, E117b.

2.2 eq NaH (60% dispersion in oil) and 2.5 eq trimethylsulfoxoniumiodide were weighed into a dry flask under nitrogen. 20 ml anhydrousDMSO was added and the reaction stirred one hour at room temperature.The intermediate acid E117b (6.8 mmol), dissolved in 6 ml anhydrous DMSOwas added dropwise. After 2.5 hours at room temperature, the reactionwas poured into 1N HCl and extracted with EtOAc. The combined organiclayers were washed with a minimal amount of aqueous sodium thiosulfatethen brine. The combined organic layers were then dried over MgSO₄,filtered and concentrated to provide the intermediate cyclopropaneE117c. The material E117c was used without further purification.

The intermediate cyclopropane (0.95 mmol) was dissolved in 5 mlanhydrous CH₂Cl₂ under nitrogen. Two drops of anhydrous DMF were addedfollowed by 1.1 eq. of oxalyl chloride. After 1.5 hours, this solutionwas added dropwise to a solution of 1.0 eq. of 4-phenoxyaniline and 1.2eq. Et₃N dissolved in 5 ml CH₂Cl₂. The reaction was left at roomtemperature overnight then poured into water. The aqueous layer wasextracted with CH₂Cl₂. The combined organic layers were dried overMgSO₄, filtered and concentrated. The intermediate amide was purified byflash chromatography.

The intermediate amide (0.125 mmol) was dissolved in 1.2 ml of 3:1THF/MeOH. 3 Eq. of LiOH.H₂O dissolved in 300 μL water was added to thissolution at 0° C. The reaction was warmed to room temperature. After onehour the reaction was quenched with sat. NH₄Cl then the pH lowered to 3with 1N HCl. The precipitate was either collected by filtration or theaqueous layer was extracted with EtOAc. The combined organic extractswere dried over MgSO₄, filtered and concentrated to give theintermediate acid. The material was used without further purification.

The intermediate acid was dissolved in 1 mL anhydrous DMF undernitrogen. 1.6 eq. EDC was added followed by 0.08 eq. DMAP and 1.3 eq.6-aminoisoquinoline and the reaction left at room temperature overnight. Reaction was poured into water and extracted with EtOAc. Thecombined organic layers were washed once with water, dried over MgSO₄,filtered and concentrated. The final compound E117d was purified byflash chromatography.

Example 118. Synthesis of4-(2-(isoquinolin-6-ylcarbamoyl)cyclopropyl)-N-(pyridin-4-yl)benzamide(E118)

Using the procedures in Example 117, Intermediate 118a was prepared. 1.2Eq. of 6-aminoisoquinoline was suspended in 0.5 ml toluene in a dryflask under nitrogen. 1.2 eq of 2.0M trimethylaluminum in heptane wasadded dropwise. After one hour all the suspended material has dissolved.This solution was added to 0.1 mmol of intermediate 118a suspended in0.5 ml toluene under nitrogen. The reaction is heated at 80° C.overnight. Sat. aq. Rochelle's salt was added to the reaction and thiswas stirred vigorously for 30 minutes. The aqueous layer was extractedwith EtOAc and the combined organic extracts were dried over MgSO₄,filtered and concentrated. The compound E118b was purified by flashchromatography.

Reference Example One The Cell-Based Porcine Trabecular Meshwork (PTM)Assay

The anterior section of porcine eyes was harvested within 4 hourspost-mortem. The iris and ciliary body were removed and trabecularmeshwork cells were harvested by blunt dissection. Finely mincedtrabecular meshwork tissue was plated into collagen-coated 6-well platesin Medium-199 containing 20% fetal bovine serum (FBS). After twopassages at confluence, cells were transferred to low-glucose DMEMcontaining 10% FBS. Cells were used between passage 3 and passage 8.

Cells were plated into fibronectin-coated, glass multiwell plates theday before compound testing under standard culture conditions. Compoundswere added to cells in the presence of 1% FBS-containing DMEM and 1%DMSO. When compounds were incubated with the cells for the durationdetermined to be optimal, the media and compound is removed and cellsfixed for 20 minutes in 3% methanol-free paraformaldehyde. Cells wererinsed twice with phosphate buffered saline (PBS) and cells arepermeabilized with 0.5% Triton X-100 for two minutes. Following anadditional two washes with PBS, F-actin was stained with Alexa-fluor488-labelled phalloidin and nuclei are stained with DAPI.

Data was reduced to the mean straight actin-fiber length and normalizedto DMSO-treated control cells (100%) and 50 μM Y-27632 (0%). Y-27632 isa rho-kinase inhibitor known to result in the depolymerization ofF-actin in these cells

Example 119. Preparation of 2-chloro-N-(5-chloro-isoquinolin-6-yl)acetamide, (E119a) and3-(2-(5-chloroisoquinolin-6-ylamino)-2-oxoethylamino)-N-methyl benzamide(E119b)

To chloroacetic acid in DMF is added EDC, DMAP and5-chloro-6-aminoisoquinoline. This mixture is stirred for 4 hours. Thereaction is washed with NaHCO₃(sat), extracted with EtOAc, dried(Na₂SO₄), filtered and evaporated. Column chromatography (SiO₂, 5%MeOH/CH₂Cl₂) gives 2-chloro-N-(5-chloroisoquinolin-6-yl) acetamide.

To 2-chloro-N-(5-chloroisoquinolin-6-yl) acetamide in MeOH is added KIand the solution is heated to 60° C. for 40 minutes. The mixture iscooled to 45° C. and 3-amino-N-methylbenzamide is added and stirred at45° C. After 2-4 hours or when TLC indicates completion of the reaction,the solvents are evaporated and the residue is taken up in EtOAc andextracted with NaHCO₃ (sat). The organics are dried (Na₂SO₄), filteredand evaporated. Flash chromatography (SiO₂, NH₃(2M) in MeOH/3%MeOH/CH₂Cl₂) gives pure3-(2-(5-chloroisoquinolin-6-ylamino)-2-oxoethylamino)-N-methylbenzamide.

Using the general procedure shown for example 119b, the followingcompounds can be synthesized from the corresponding 6-aminoisoquinoline.

Example R R₂ R₁ 120 3-CONHMe 5-Cl H 121 3-CONHMe 5-Br H 122 3-CONHMe5-Me H 123 3-CONHMe 5-nPr H 124 3-CONHMe 5-CH═CH₂ H 125 3-CONHMe5-CH₂CH═CH₂ H 126 3-CONHMe 5,8-diMe H 127 3-CONHMe 7-Me 1-Cl 128 3-COMe5-Cl H 129 3-COMe 5-Me H 130 3-CO₂Me 5-Cl H 131 3-CO₂Me 5-Me H 132 3-Cl5-Cl H 133 3-Cl 5-Me H 134 3-OMe 5-Cl H 135 3-OMe 5-Me H

Example 136. Preparation of3-((N-isoquinolin-6-ylsulfamoyl)methylamino)-N-methylbenzamide (E136)

To 6-aminoisoquinoline in DMF at 0° C. is added NaH. After 30 min,chlorosulfonyl chloride is added to the reaction. After 2-4 hours at rtor when TLC indicates completion, the reaction is quenched by theaddition of water and extracted with EtOAc. The combined organics arewashed with brine and dried (Na₂SO₄), filtered and evaporated. Columnchromatography (SiO₂, 5% MeOH/CH₂Cl₂) gives1-chloro-N-(isoquinolin-6-yl) methanesulfonamide.

To 1-chloro-N-(isoquinolin-6-yl) methanesulfonamide in MeOH is added KIand the solution is heated to 60° C. for 40 minutes. The mixture iscooled to 45° C. and 3-amino-N-methylbenzamide is added and stirred at45° C. After 2-4 hours or when TLC indicated completion of the reaction,the solvents are evaporated and the residue is taken up in EtOAc andextracted with NaHCO₃ (sat). The organics are dried (Na₂SO₄), filteredand evaporated. Flash chromatography (SiO₂, 2% NH₃(2M) in MeOH/3%MeOH/CH₂Cl₂) gives3-((N-isoquinolin-6-ylsulfamoyl)methylamino)-N-methylbenzamide.

Examples 137-141

Using the general procedure shown for example 136, the followingcompounds can be synthesized from the corresponding 6-aminoisoquinoline.

Example R 137 3-CONH₂ 138 3-COMe 139 3-CO₂Me 140 3-Cl 141 3-OMe

Example 142

Using the general procedure shown for Example 119, the followingcompound was synthesized from the corresponding cycloalkylamine.

Example No. n R 142 2 H

Examples 143-147

Using the general procedure shown for Example 119, the followingcompounds could be synthesized from the corresponding cycloalkylamines.

Example No. n R 143 1 H 144 2 3-CONHMe 145 2 3-CO₂Me 146 2 3-COMe 147 23-Cl

Example 148. Preparation2-(4-(1-(4-chlorobenzyl)-1H-1,2,3-triazol-4-yl)phenoxy)-N-(isoquinolin-6-yl)-acetamide(E148)

A solution of 4-acetamidobenzenesulfonyl azide in acetonitrile wastreated with cesium carbonate and stirred at room temperature for 5minutes. Dimethyl 2-oxopropylphosphonate was added and stirred for 1hour. Then, a solution of methyl 2-(4-formylphenoxy)acetate in MeOH wasadded to the reaction and stirred for an additional 2 hours. Thesolvents were removed and the residue was dissolved in diethyl ether andwashed with water and brine. The organics were dried (Na₂SO₄), filteredand concentrated. Flash chromatography (SiO₂, EtOAc/hexanes) gave thecorresponding alkyne.

A solution of 4-chlorobenzyl bromide in acetone was treated with NaN₃and refluxed for 3 hours. The reactions was concentrated by 50% anddiluted with saturated sodium chloride solution. The reaction was thenextracted with Et₂O, washed with brine, dried (Na₂SO₄) and concentratedto give 1-(azidomethyl)-4-chlorobenzene.

A 5 mL microwave reaction vial was charged with methyl2-(4-ethynylphenoxy)acetate, 1-(azidomethyl)-4-chlorobenzene, t-BuOH,copper turnings and copper sulfate. The reaction was heated undermicrowave conditions at 125° C. for 25 minutes. The reaction was cooledto room temperature and poured into water. The reaction was extractedwith CH₂Cl₂. The combined organic layers were washed with water, dried(Na₂SO₄), filtered and concentrated. Flash chromatography (SiO₂,Hexanes/EtOAc) gave2-(4-(1-(4-chlorobenzyl)-1H-1,2,3-triazol-4-yl)phenoxy)-N-(isoquinolin-6-yl)-acetamide(E148).

Example 149. Preparation of2-(4-(1-benzyl-1H-tetrazol-5-yl)phenoxy-N-(isoquinolin-6-yl)acetamide(E149)

A mixture of 4-cyanophenol and methyl bromoacetate in CH₃CN was treatedwith K₂CO₃ and refluxed for 18 hours. Then the reaction was cooled toroom temperature and concentrated. The residue was partitioned betweenEtOAc and H₂O and the organic layer was washed with brine, driend(Na₂SO₄), filtered and evaporated. Flash chromatography (SiO₂,hexanes/EtOAc) gave pure methyl 2-(4-cyanophenoxy)acetate.

A solution methyl 2-(4-cyanophenoxy)acetate in DMF was treated with NaN₃and NH₄Cl and stirred in an oil bath at 80° C. for 18 hours. Then, themixture was poured into H₂O and extracted with Et₂O. The aqueous layerwas acidified to pH 2-3 with HCl and the precipitate was collected togive methyl 2-(4-(2H-tetrazol-5-yl)phenoxy)acetate.

A solution of methyl 2-(4-(2H-tetrazol-5-yl)phenoxy)acetate in acetonewas treated with triethylamine and stirred at room temperature for 20minutes. Benzyl bromide was added and stirred for 18 hours at 60° C. Thesolvent was removed and the residue was purified by reverse phasepreparative HPLC to give2-(4-(1-(4-chlorobenzyl)-1H-1,2,3-triazol-4-yl)phenoxy)-N-(isoquinolin-6-yl)-acetamide(E149).

Using largely the procedures set forth in Examples 148-149 andsubstituting the appropriate starting material, the compounds 150-153were made.

Example No. A X 150

m-CH₂—C₆H₄—F 151

m-CH₂—C₆H₄—OMe 152

o-CH₂—C₆H₄—CH₃ 153

m-CH₂—C₆H₄—OMe

Example 154

Using largely the procedure set forth in Examples 117 and 118 andsubstituting the appropriate starting materials the compounds 154-157were made.

Example Ar 154 Ph— 155

156

157

Example 158-169

Using essentially the procedure set forth in Examples 117 and 118 andsubstituting the appropriate starting materials the compounds 158-169can be made.

Example Ar 158 m-F—C₆H₄ 159 m-MeO—C₆H₄ 160 m-COOMe—C₆H₄ 161 2-pyridyl162 3-pyridyl 163 4-pyridyl 164

165

166

167

Example 168. Synthesis of(1S,2S)-N-(isoquinolin-6-yl)-2-phenylcyclopropane carboxamide: (E168)

The title compound is obtained as described in Example 117.

Example 169. Synthesis of(1R,2R)-N-(isoquinolin-6-yl)-2-phenylcyclopropane carboxamide (E169)

The title compound is obtained as described in Example 117.

The chiral non-racemic materials described in these examples may beobtained by following the references set herein: Resolution oftrans-phenylcyclopropane carboxylic acids via the quinine salt: Inouye,Y.; Sugita, T.; Walborsky, H. M. Tetrahedron 1964, 20, 1695; Webster, F.X.; Zeng, X.; Silverstein, R. M. J. Org. Chem. 1982, 47, 5225, andreferences cited therein. Reference for resolution oftrans-phenylcyclopropane carboxylic acids via separation on a chiralHPLC column: Penmetsa, K. V.; Reddick, C. D.; Fink, S. W.; Kleintop, B.L.; DiDonato, G. C.; Volk, K. J.; Klohr, S. E. J. Liq. Chrom. & Rel.Technol. 2000, 23, 831, and references cited therein.

Examples 170-178

Using largely the procedure set forth in Examples 117 and 118 butreplacing the cyclopropanation with a hydrogenation and substituting theappropriate starting materials the compounds 170-178 were made.

Example Ar 170 o-NH₂Ph— 171

172

173

174

175

176

177

178

Example 179-186

Using largely the procedure set forth in Examples 117 and 118 butreplacing the cyclopropanation with a hydrogenation and substituting theappropriate starting materials the compounds 179-186 can be made.

Example Ar 179 p-F—C₆H₄ 180 m-CN—C₆H₄ 181 m-CF₃—C₆H₄ 182 m-NMe₂—C₆H₄ 183o-F—C₆H₄ 184 p-SMe—C₆H₄ 185

186

Example 187-188

Using largely the procedure set forth in Examples 117 and 118 buteliminating the cyclopropanation step and substituting the appropriatestarting materials the compounds 187 and 188 were made.

Example Ar 187 meta-NH₂Ph— 188

Examples 189-198

Using largely the procedure set forth in Examples 117 and 118 buteliminating the cyclopropanation step and substituting the appropriatestarting materials the compounds 189-198 can be made.

Example Ar 189 m-Cl—C₆H₄ 190 p-COOMe—C₆H₄ 191 2-furan 192 3-furan 1932-thiophene 194 3-thiophene 195 p-COMe—C₆H₄ 196 m-NO₂—C₆H₄ 197m-NH₂—C₆H₄ 198 m-OAc—C₆H₄

Examples 199-204

Using largely the procedure set forth in Examples 117 and 118 andsubstituting the appropriate starting materials the compounds 199-204can be made.

Example Ar B 199 Ph—

200 Ph—

201 Ph— —C≡C— 202 Ph— C≡C—CH₂ 203 Ph— trans-CH₂—CH═CH— 204 Ph—trans-CH═CH—CH₂—

Example 205. Synthesis of(E)-N-(isoquinolin-6-yl)-3-(4-(2-phenylacryloyl)phenyl) acrylamide(E205)

A round bottomed flask equipped with a condenser and a stirring bar wascharged with benzyl 4-bromophenylketone (27 mmol), sodium acetate (1.2eq) tert-butyl acrylate (1.2 eq) and N-methylpyrrolidinone (NMP) (35ml). In a separate flask, 22.5 mg Pd(OAc)₂ was dissolved in 50 ml NMP.6.75 ml of this solution was added to the reaction (0.05 mol %). Thereaction mixture was heated to 135° C. for 40 minutes. After cooling toroom temperature, the reaction was quenched with water then extractedwith EtOAc. The combined organic layers were washed with water and brineand dried over Na₂SO₄. This solution was filtered though celite toremoved the catalyst then concentrated to give the ester intermediateE205b. The material was used without further purification.

The ester intermediate E205b was dissolved in acetic anhydride (4 eq.)then N,N,N′,N′-tetramethyldiaminomethane (2.5 eq) was added dropwise viasyringe. After 40 minutes sat. NaHCO₃ was added and the resultingmixture extracted with EtOAc. The combined organic layers were washedwith sat. NaHCO₃, 1N HCl and brine, dried over Na₂SO₄ and concentrated.The compound was purified by column chromatography to give the alkeneintermediate E205c.

The alkene intermediate E205c (13.8 mmol) was dissolved in DCM andcooled to 0° C. 15 mL Trifluoroacetic acid was added and the reactionstirred at 0° C. for 1 hour then room temperature for 2 hours. Thesolvents were evaporated and the solid residue suspended in ether.Filtration gave the intermediate acid.

The intermediate acid (0.14 mmol) was dissolved in 0.8 ml anhydrous DMFunder nitrogen. 1.6 Eq. EDC was added followed by 0.08 eq. DMAP and 1.3eq. 6-aminoisoquinoline and the reaction left at room temperature for 3hours. The reaction was poured into water and extracted with EtOAc. Thecombined organic layers were washed once with water, dried over Na₂SO₄,filtered and concentrated. The compound was purified by flashchromatography.

Example 206. Synthesis of(E)-N-(isoquinolin-6-yl)-3-(4-(1-phenylcyclopropane carbonyl)phenyl)acrylamide (E206)

Using the procedure set forth in Example 205 intermediate E206a wasmade.

2.2 eq NaH (60% dispersion in oil) and 2.5 eq trimethylsulfoxoniumiodide were weighed into a dry flask under nitrogen. Anhydrous DMSO wasadded and the reaction stirred one hour at room temperature. Theintermediate acid E206a (0.36 mmol), dissolved in anhydrous DMSO wasadded dropwise. After 5 minutes at room temperature, the reaction waspoured into 1N HCl and extracted with EtOAc. The combined organic layerswere washed with a minimal amount of aqueous sodium thiosulfate thenbrine. The combined organic layers were then dried over MgSO₄, filteredand concentrated to provide the intermediate cyclopropane. The materialwas purified by flash chromatography to give the intermediate acid.

The intermediate acid (0.12 mmol) was dissolved in anhydrous DMF undernitrogen. 1.6 eq. EDC was added followed by 0.08 eq. DMAP and 1.3 eq.6-aminoisoquinoline and the reaction left at room temperature for 3hours. Reaction was poured into water and extracted with EtOAc. Thecombined organic layers were washed once with water, dried over MgSO₄,filtered and concentrated. The compound E206b was purified by flashchromatography.

Example 207. Synthesis of(2R)-2-amino-3-(3-(4-((E)-3-(isoquinolin-6-ylamino)-3-oxoprop-1-enyl)phenyl)-3-oxo-2-phenylpropylthio)propanoicacid (E207)

Intermediate E207a and (L)-cystine were suspended in 2:1 phosphatebuffer/ACN. NaOH was added and the reaction stirred at room temperature.When reaction was complete by LC/MS, the pH was adjusted to 6. Theprecipitate that was formed was collected by filtration.

Example 208. Synthesis ofN-(isoquinolin-6-yl)-3-(4-(3-phenylpropanamido)phenyl) propanamide(E208b)

A solution of 3-(4-aminophenyl)propionic acid in methanol was cooled to0° C. Thionyl chloride was added dropwise. Following addition, thereaction was heated to 40° C. overnight. The reaction was concentratedto provide the intermediate methyl ester which was used without furtherpurification.

The intermediate ester (0.115) was dissolved in anhydrous THF undernitrogen. EDC was added followed by DMAP and hydrocinnamic acid. Thereaction was left at room temperature over night. The reaction waspoured into water and extracted with EtOAc. The combined organic layerswere washed once with water, dried over MgSO₄, filtered andconcentrated. The compound was purified by flash chromatography to givethe intermediate amide (E208a).

The intermediate amide (E208a) was dissolved in 3:1 THF/MeOH. LiOH.H2Odissolved in water was added to this solution at 0° C. The reaction waswarmed to room temperature. After one hour the reaction was quenchedwith sat. NH₄Cl then the pH lowered to 3 with 1N HCl. The precipitatewas collected by filtration or the aqueous layer was extracted withEtOAc. The combined organic extracts were dried over MgSO₄, filtered andconcentrated to give the intermediate acid. The material was usedwithout further purification.

The intermediate acid was dissolved in anhydrous DMF under nitrogen. 1.6eq. EDC was added followed by 0.08 eq. DMAP and 1.3 eq.6-aminoisoquinoline and the reaction left at room temperature overnight. Reaction was poured into water and extracted with EtOAc. Thecombined organic layers were washed once with water, dried over MgSO₄,filtered and concentrated. The compound (E208b) was purified by flashchromatography.

Example 209. Synthesis ofN-(4-(3-(isoquinolin-6-ylamino)-3-oxopropyl)phenyl)-4-methoxybenzamide(E209)

The title compound was obtained as described in Example 208

Example 210. Synthesis ofN-(isoquinolin-6-yl)-3-(4-(3-(3-(trifluoromethyl)phenyl)ureido)phenyl)propanamide (E110)

Steps one, three and four were performed as in Example 208. Step two isperformed as follows: To methyl 3-(4-aminophenyl)propanoate in DMF isadded DMAP and 3-trifluoromethylphenyl isocyanate and the solution wasstirred at room temperature for 4 hours. The mixture was poured intoNaHCO₃ (sat), extracted with EtOAc, dried (Na₂SO₄), filtered andevaporated. The compound E210b was purified by flash chromatography.

Example 211. Synthesis of(E)-3-(4-(3-(2-fluoro-5-methylphenyl)ureido)phenyl)-N-(isoquinolin-6-yl)acrylamide(E211)

The title compound was obtained as described in Example 210.

Reference Example Two Pharmacological Activity for Glaucoma Assay

Pharmacological activity for glaucoma can be demonstrated using assaysdesigned to test the ability of the subject compounds to decreaseintraocular pressure. Examples of such assays are described in thefollowing reference, incorporated herein by reference: C. Liljebris, G.Selen, B. Resul, J. Sternschantz, and U. Hacksell, “Derivatives of17-Phenyl-18, 19, 20-trinorprostaglandin F_(2α) Isopropyl Ester:Potential Anti-glaucoma Agents”, Journal of Medicinal Chemistry, Vol. 38(2) 1995, pp. 289-304.

Example 212

Topical pharmaceutical compositions for lowering intraocular pressureare prepared by conventional methods and formulated as follows:

Ingredient Amount (wt %) 6-aminoisoquinoline Derivative 0.50 Dextran 700.1 Hydroxypropyl methylcellulose 0.3 Sodium Chloride 0.77 Potassiumchloride 0.12 Disodium EDTA 0.05 Benzalkonium chloride 0.01 HCl and/orNaOH pH 7.0-7.2 Purified water q.s. to 100%

A compound according to this invention is used as the6-aminoisoquinoline derivative. When the composition is topicallyadministered to the eyes once daily, the above composition decreasesintraocular pressure in a patient suffering from glaucoma.

Example 213

Example 212 is repeated using(2R)-2-amino-3-(3-(4-((E)-3-(isoquinolin-6-ylamino)-3-oxoprop-1-enyl)phenyl)-3-oxo-2-phenylpropylthio) propanoic acid (E207) according to this invention. Whenadministered as a drop 4 times per day, the above compositionsubstantially decreases intraocular pressure and serves as aneuroprotective agent.

Example 214

Example 212 is repeated using2-(benzylamino)-N-(isoquinolin-6-yl)acetamide (E58) according to thisinvention. When administered as a drop twice per day, the abovecomposition substantially decreases intraocular pressure.

Example 215

Example 212 is repeated using 2-chloro-N-(isoquinolin-6-yl) acetamide(E1) according to this invention. When administered as a drop twice perday, the above composition substantially decreases allergic symptoms andrelieves dry eye syndrome.

Example 216

Example 212 is repeated using 2-amino-N-isoquinoline-6-yl-acetamidedihydrochloride (E40) according to this invention. When administered asa drop as needed, the above composition substantially decreaseshyperemia, redness and ocular irritation.

Example 217

Example 212 is repeated usingN-(isoquinolin-6-yl)-2-(3-methoxybenzylamino)acetamide (E59) accordingto this invention. When administered as a drop 4 times per day, theabove composition substantially decreases intraocular pressure andserves as a neuroprotective agent.

Example 218

Example 212 is repeated using3-((N-isoquinolin-6-ylsulfamoyl)methylamino)-N-methylbenzamide (E136)according to this invention. When administered as a drop twice per day,the above composition substantially decreases intraocular pressure.

Example 219

Example 212 is repeated using benzylisoquinolin-6-ylcarbamate (E87).according to this invention. When administered as a drop twice per day,the above composition substantially decreases allergic symptoms andrelieves dry eye syndrome.

Example 220

Example 213 is repeated using 1-benzyl-3-(isoquinolin-6-yl)urea (E88)according to this invention. When administered as a drop as needed, theabove composition substantially decreases allergic symptoms

Example 221

Example 213 is repeated using N-(isoquinolin-6-yl)-2-morpholinoacetamide(E105) according to this invention. When administered as a drop asneeded, the above composition substantially decreases hyperemia, rednessand ocular irritation.

Example 222

Example 213 is repeated using4-(2-(isoquinolin-6-ylcarbamoyl)cyclopropyl)-N-(pyridin-4-yl)benzamide(E118) according to this invention. When administered as a drop twice aday or as needed, the above composition substantially decreasesintraocular pressure.

Example 223

Example 213 is repeated using3-(2-(5-chloroisoquinolin-6-ylamino)-2-oxoethylamino)-N-methylbenzamide(E119b) according to this invention. When administered as a drop twice aday or as needed, the above composition substantially decreasesintraocular pressure.

While the invention has been described in detail and with reference tospecific embodiments thereof, it will be apparent to one skilled in theart that various changes and modifications can be made without departingfrom the spirit and scope of the invention.

What is claimed is:
 1. A compound, wherein the compound is:

or a pharmaceutically acceptable salt thereof.
 2. A method ofinfluencing the action of a kinase in a cell, a tissue, or a livingmammal comprising administering to or contacting with the cell, tissue,or mammal at least one compound according to claim 1, or increasing theeffectiveness of another therapeutic agent in a cell, tissue or livingmammal comprising administering to or contacting with the cell, tissueor mammal a therapeutically effective amount of at least one compound ofclaim
 1. 3. The method of claim 2, wherein the kinase affected is atyrosine kinase.
 4. The method of claim 2, wherein the kinase affectedis ROK-I, ROK-II, PKA, PKC, a CAM kinase, GRK-2, GRK-3, GRK-5 or GRK-6.5. A pharmaceutical composition comprising a compound of claim 1 and acarrier.
 6. The composition of claim 5, wherein the carrier is asystemic carrier or a topical carrier.
 7. The composition of claim 5,wherein the composition comprises about 0.001% to 10% of the compoundand 90 to 99.999% of the carrier, wherein the carrier is a systemiccarrier.
 8. A method of treating a condition in a subject in needthereof, comprising administering a safe and effective amount of acompound of claim 1 to the subject, wherein the condition is an eyedisease, a bone disorder, obesity, a heart disease, a hepatic disease, arenal disease, pancreatitis, a cancer, myocardial infarct, gastricdisturbance, hypertension, fertility control, nasal congestion,neurogenic bladder disorder, a gastrointestinal disorder, or adermatological disorder.
 9. The method of claim 8, wherein the conditionis an eye disease.
 10. The method of claim 8, wherein the condition isglaucoma.
 11. The method of claim 8, wherein the condition is aneurodegenerative eye disease.